Information on EC 3.4.13.20 - beta-Ala-His dipeptidase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.4.13.20
-
RECOMMENDED NAME
GeneOntology No.
beta-Ala-His dipeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
preferential hydrolysis of the beta-Ala-/-His dipeptide (carnosine), and also anserine, Xaa-/-His dipeptides and other dipeptides including homocarnosine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Arginine and proline metabolism
-
-
beta-Alanine metabolism
-
-
Histidine metabolism
-
-
Metabolic pathways
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
9027-21-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain MCI3434
UniProt
Manually annotated by BRENDA team
strain MCI3434
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
HeLa-CN1 cells have a lower cell survival than do non-transfected cells or HeLa cells transfected with an empty or a mock vector. Viability in HeLa cells measured after H2O2 treatment is significantly higher than the viability of HeLa-CN1 cells after the same oxidant insult, indicating an increased susceptibility to oxidative stress of cells overexpressing carnosinase
metabolism
-
carnosine and anserine are mainly hydrolyzed by carnosinase, a low but significant amount is excreted in the urine. Carnosine reaches a steady state of very low concentration in serum, while anserine sustains higher concentrations than that of carnosine due to its greater stability vis-a-vis carnosinase after ingesting histidine-dipeptide rich diet
physiological function
-
CN1 activity is lower in children and increases with age with no differences in protein concentration. CN1 activity is considerably higher in serum than in cerebrospinal fluid
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(beta-homoAla-beta-homoLys-beta-homoPhe)2 + H2O
beta-homoAla + beta-homoLys-beta-homoPhe-beta-homoAla-beta-homoLys-beta-homoPhe
show the reaction diagram
3-(acetylamino)-L-alanyl-L-histidine + H2O
3-(acetylamino)-L-alanine + L-histidine
show the reaction diagram
-
3% of the activity with carnosine
-
-
?
3-amino-L-alanyl-L-histidine + H2O
3-amino-L-alanine + L-histidine
show the reaction diagram
-
30% of the activity with carnosine
-
-
?
Ala-Ala + H2O
Ala + Ala
show the reaction diagram
-
11% of the activity with carnosine
-
-
?
Ala-His + H2O
Ala + His
show the reaction diagram
-
38% of the activity with carnosine
-
-
?
Ala-Leu + H2O
Ala + Leu
show the reaction diagram
-
13% of the activity with carnosine
-
-
?
Ala-Tyr + H2O
Ala + Tyr
show the reaction diagram
-
8% of the activity with carnosine
-
-
?
anserine + H2O
beta-Ala + Ntau-methyl-L-histidine
show the reaction diagram
beta-Ala-4-nitroanilide + H2O
beta-Ala + 4-nitroaniline
show the reaction diagram
beta-Ala-Ala + H2O
beta-Ala + Ala
show the reaction diagram
-
10% of the activity with carnosine
-
-
?
beta-Ala-beta-Ala + H2O
beta-Ala + beta-Ala
show the reaction diagram
beta-Ala-Gly + H2O
beta-Ala + Gly
show the reaction diagram
76% of the activity compared to beta-Ala-L-Ala
-
-
?
beta-Ala-His + H2O
beta-Ala + His
show the reaction diagram
-
i.e. carnosine
-
-
?
beta-Ala-Ile-beta-homoTyr + H2O
beta-Ala + Ile-beta-homoTyr
show the reaction diagram
beta-Ala-L-Ala + H2O
beta-Ala + L-Ala
show the reaction diagram
preferred substrate
-
-
?
beta-Ala-L-His + H2O
beta-Ala + L-His
show the reaction diagram
beta-Ala-L-Leu + H2O
beta-Ala + L-Leu
show the reaction diagram
49% of the activity compared to beta-Ala-L-Ala
-
-
?
beta-Ala-NH2 + H2O
beta-Ala + NH3
show the reaction diagram
58% of the activity compared to beta-Ala-L-Ala
-
-
?
beta-Ala-Phe + H2O
beta-Ala + Phe
show the reaction diagram
-
18% of the activity with carnosine
-
-
?
beta-homoAla-4-nitroanilide + H2O
beta-homoAla + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
beta-homoAla-beta-homoLeu + H2O
beta-homoAla + beta-homoLeu
show the reaction diagram
-
hydrolysis at 55% compared to hydrolysis of carnosine
-
-
?
beta-homoLeu-Ile-beta-homoTyr + H2O
beta-homoLeu + Ile-beta-homoTyr
show the reaction diagram
-
hydrolysis at 0.01% compared to hydrolysis of carnosine
-
-
?
beta-homoPhe-4-nitroanilide + H2O
beta-homoPhe + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
beta-homoSer-Ile-beta-homoTyr + H2O
beta-homoSer + Ile-beta-homoTyr
show the reaction diagram
-
hydrolysis at 0.06% compared to hydrolysis of carnosine
-
-
?
beta-homoVal-beta-homoAla-beta-homoLeu + H2O
beta-homoVal + beta-homoAla-beta-homoLeu
show the reaction diagram
-
hydrolysis at 0.19% compared to hydrolysis of carnosine
-
-
?
beta-homoVal-Ile-beta-homoTyr + H2O
beta-homoVal + Ile-beta-homoTyr
show the reaction diagram
-
hydrolysis at 0.01% compared to hydrolysis of carnosine
-
-
?
beta-homoVal-Ile-Tyr + H2O
beta-homoVal + Ile-Tyr
show the reaction diagram
-
hydrolysis at 0.09% compared to hydrolysis of carnosine
-
-
?
carnosine + H2O
?
show the reaction diagram
-
-
-
-
?
carnosine + H2O
beta-Ala + His
show the reaction diagram
D-Ala-4-nitroanilide + H2O
D-Ala + 4-nitroaniline
show the reaction diagram
D-Ala-NH2 + H2O
D-Ala + NH3
show the reaction diagram
0.6% of the activity compared to beta-Ala-L-Ala
-
-
?
Gly-4-nitroanilide + H2O
Gly + 4-nitroaniline
show the reaction diagram
Gly-Gly + H2O
Gly + Gly
show the reaction diagram
-
7% of the activity with carnosine
-
-
?
Gly-His + H2O
Gly + His
show the reaction diagram
-
37% of the activity with carnosine
-
-
?
Gly-His-Gly + H2O
?
show the reaction diagram
-
13% of the activity with carnosine
-
-
?
Gly-His-Lys + H2O
?
show the reaction diagram
-
7% of the activity with carnosine
-
-
?
Gly-L-His + H2O
Gly + L-His
show the reaction diagram
-
-
-
?
Gly-Leu + H2O
Gly + Leu
show the reaction diagram
-
21% of the activity with carnosine
-
-
?
homocarnosine + H2O
?
show the reaction diagram
L-Ala-4-nitroanilide + H2O
L-Ala + 4-nitroaniline
show the reaction diagram
L-Ala-L-His + H2O
L-Ala + L-His
show the reaction diagram
-
-
-
?
L-Arg-4-nitroanilide + H2O
L-Arg + 4-nitroaniline
show the reaction diagram
L-carnosine + H2O
beta-Ala + His
show the reaction diagram
L-Lys-4-nitroanilide + H2O
L-Lys + 4-nitroaniline
show the reaction diagram
L-Phe-4-nitroanilide + H2O
L-Phe + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Leu-Leu + H2O
Leu + Leu
show the reaction diagram
-
8% of the activity with carnosine
-
-
?
N-acetyl-3-(acetylamino)-L-alanyl-L-histidine + H2O
N-acetyl-3-(acetylamino)-L-alanine + L-histidine
show the reaction diagram
-
63% of the activity with carnosine
-
-
?
N-methylcarnosine + H2O
N-methyl-beta-Ala + L-His
show the reaction diagram
-
-
-
?
Phe-Ala + H2O
Phe + Ala
show the reaction diagram
-
8% of the activity with carnosine
-
-
?
Ser-His + H2O
Ser + His
show the reaction diagram
-
8% of the activity with carnosine
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
anserine + H2O
beta-Ala + Ntau-methyl-L-histidine
show the reaction diagram
-
splitting in the blood stream
-
-
?
carnosine + H2O
beta-Ala + His
show the reaction diagram
-
splitting in the blood stream
-
?
homocarnosine + H2O
?
show the reaction diagram
-
splitting of homocarnosine in the brain
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Zn2+ is not required for enzymatic activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-(acetylamino)-L-alanyl-L-histidine
-
2.3 mM, 50% inhibition of 3-nitrotyrosine formation
3-amino-L-alanyl-L-histidine
-
3.3 mM, 50% inhibition of 3-nitrotyrosine formation
AgNO3
30°C, 10 min, 1 mM, 95% loss of activity
bestatin
CdCl2
30°C, 10 min, 1 mM, 90% loss of activity
citrate
-
citrate ions are shown to bind at only three well-defined sites involving both ion pairs and hydrogen bonds. Molecular dynamics simulations evidence that citrate binding has a remarkable conformational influence on the 3D structure of carnosinase, increasing the binding affinity of carnosine to the catalytic site
dithiothreitol
30°C, 10 min, 1 mM, 63% loss of activity
EDTA
-
-
HgCl2
30°C, 10 min, 1 mM, 99% loss of activity
homocarnosine
-
-
N-acetyl-3-(acetylamino)-L-alanyl-L-histidine
-
2.5 mM, 50% inhibition of 3-nitrotyrosine formation
N-ethylmaleimide
30°C, 10 min, 1 mM, 80% loss of activity
p-chloromercuribenzoate
30°C, 10 min, 1 mM, 95% loss of activity
ZnCl2
30°C, 10 min, 1 mM, 98% loss of activity
ZnSO4
30°C, 10 min, 1 mM, 98% loss of activity
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
citrate
-
activates
phosphate
-
activates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0039
beta-Ala-4-nitroanilide
-
pH 8.0, 25°C
1.27 - 11
beta-Ala-L-His
0.019
beta-homoAla-4-nitroanilide
-
pH 8.0, 25°C
1.1
beta-homoPhe-4-nitroanilide
-
pH 8.0, 25°C
0.175 - 0.21
carnosine
0.52 - 0.54
D-Ala-4-nitroanilide
3
Gly-4-nitroanilide
-
100 mM Tris/HCl or 50 mM potassium phosphate, pH 8.0, 30°C
0.2 - 1.9
homocarnosine
0.36 - 0.6
L-Ala-4-nitroanilide
0.4
L-Arg-4-nitroanilide
-
50 mM potassium phosphate, pH 8.0, 30°C
0.4
L-Lys-4-nitroanilide
-
50 mM potassium phosphate, pH 8.0, 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
30.5
beta-Ala-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
10.6 - 138
beta-Ala-L-His
51.4
beta-homoAla-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
0.03
beta-homoPhe-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
3.3 - 4
D-Ala-4-nitroanilide
70
Gly-4-nitroanilide
Ochrobactrum anthropi
-
100 mM Tris/HCl or 50 mM potassium phosphate, pH 8.0, 30°C
0.2 - 18
homocarnosine
0.5 - 0.56
L-Ala-4-nitroanilide
0.14
L-Arg-4-nitroanilide
Ochrobactrum anthropi
-
50 mM potassium phosphate, pH 8.0, 30°C
0.11
L-Lys-4-nitroanilide
Ochrobactrum anthropi
-
50 mM potassium phosphate, pH 8.0, 30°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7800
beta-Ala-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
27611
2600
beta-homoAla-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
40526
0.027
beta-homoPhe-4-nitroanilide
Ochrobactrum anthropi
-
pH 8.0, 25°C
40527
6.3 - 7.5
D-Ala-4-nitroanilide
5809
23
Gly-4-nitroanilide
Ochrobactrum anthropi
-
100 mM Tris/HCl or 0 mM potassium phosphate, pH 8.0, 30°C
9764
0.8 - 1.55
L-Ala-4-nitroanilide
1891
0.35
L-Arg-4-nitroanilide
Ochrobactrum anthropi
-
50 mM potassium phosphate, pH 8.0, 30°C
8550
0.275
L-Lys-4-nitroanilide
Ochrobactrum anthropi
-
50 mM potassium phosphate, pH 8.0, 30°C
3734
0.017
L-Phe-4-nitroanilide
Ochrobactrum anthropi
-
50 mM potassium phosphate, pH 8.0, 30°C
7222
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000007
bestatin
-
pH 7.5, 30°C
0.24
homocarnosine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
assay at
7.5 - 8.5
-
broad
8 - 8.5
-
-
8.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
CN1 activity is considerably higher in serum than in cerebrospinal fluid
Manually annotated by BRENDA team
-
low expresion
Manually annotated by BRENDA team
-
highly expressed in the mitral cell layer of the olfactory bulb
Manually annotated by BRENDA team
-
highly expressed in parafascicular nucleus of the thalamus
Manually annotated by BRENDA team
additional information
-
the enzyme activity in tissues is roughly proportional to blood content for all the tissues except brain, indicating that serum carnosinase is present in the trapped blood of these tissues
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Haemophilus somnus (strain 129Pt)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
13000
alpha4beta4, 2 * 13000 + 2 * 27000, SDS-PAGE
13737
-
x * 13737 + x * 26564, electrospray mass spectrometry
15000
-
x * 15000 + x * 30000, SDS-PAGE
26564
-
x * 13737 + x * 26564, electrospray mass spectrometry
27000
alpha4beta4, 2 * 13000 + 2 * 27000, SDS-PAGE
30000
-
x * 15000 + x * 30000, SDS-PAGE
52800
-
2 * 82000, SDS-PAGE, 2 * 63700, MALDI-TOF, 2 * 52800, calculated
63700
-
2 * 82000, SDS-PAGE, 2 * 63700, MALDI-TOF, 2 * 52800, calculated
65000
-
SDS-PAGE, non-reducing conditions
75000
-
2 * 75000, subunits are connected by one or more disulfide bonds, SDS-PAGE
82000
-
2 * 82000, SDS-PAGE, 2 * 63700, MALDI-TOF, 2 * 52800, calculated
86000
-
x* 92000, SDS-PAGE of secreted protein, x * 86000 and x * 88000, SDS-PAGE of cytosolic protein
88000
-
x* 92000, SDS-PAGE of secreted protein, x * 86000 and x * 88000, SDS-PAGE of cytosolic protein
130000
-
SDS-PAGE, reducing conditions
150000
160000
-
gel filtration
167000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
x-ray crystallography
monomer
-
1 * 65000, SDS-PAGE
octamer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
complexed with bestatin together with Zn2+, hanging drop vapour diffusion method, using 20% polyethylene glycol 3350 and 0.2 M potassium fluoride
-
active-site modeling of DmpA. Based on the crystal structure and the catalytic mechanism proposed for DmpA, carnosine is manually introduced into the active site of the enzyme. The model shows that carnosine fits excellently, in line with the high specific activity for this substrate
-
crystal structure of the enzyme determined to 1.82 A resolution using the multiple isomorphous replacement method. The heterodimer folds into a single domain organised as an alphabetabetaalpha sandwich in which two mixed beta sheets are flanked on both sides by two alpha helices
-
hangig-drop vapor-diffusion method. Two crystal forms are obtained at 21°C in 13-16% PEG 2000 monomethylether at pH 9.0, adding either 0.2 M magnesium chloride or 1 M lithium chloride. Crystals of the first form belong to the space group C222(1) and diffract to 3.0 A resolution. Crystals of the second form belong to the space group P2(1)2(1)2 and diffract to 2.3 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 11
30°C, 10 min, stable
708466
7.5 - 8.2
-
maximal stability at 50°C
647115
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
up to 8 h the activity remains within 97.5% of the initial value, thereafter the activity decreases slowly
45
10 min, stable below
55
10 min, 51% loss of activity
60
10 min, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
inactivation during DEAE-cellulose chromatography, slow reactivation when concentrated and stored at 4°C
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, stable for several weeks in serum
-
4°C, stable for a day or two in serum
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
glutathione-Sepharose bead chromatography and Superdex 200 gel filtration
-
recombinant enzyme
-
recombinant enzyme is purified from the Escherichia coli JM109 harboring p2DAPEX with a recovery of 10.3%
using DEAE Sephacel and UnoQ chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a stable HeLa cell line expressing recombinant human serum carnosinase CN1 is established
-
expressed in Escherichia coli strain BL21(DE3)pLysS
-
expression in CHO cell
-
expression in COS-7 cell
-
expression in db/db mice, a model of type 2 diabetes
-
expression in Escherichia coli
human CN1 gene is expressed in Saccharomyces cerevisiae on the yeast cell surface with alpha-agglutinin as the anchor protein
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D166A
-
mutation in putative metal binding site, complete loss of activity
E201A
-
mutation in putative metal binding site, complete loss of activity
E201A/D229A
-
mutation in putative metal binding site, complete loss of activity
H133A
-
mutation in putative metal binding site, complete loss of activity
H228A
-
inactive
G249A
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
G249D
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
S250A
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
S250C
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
S250T
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
G249A
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
-
G249D
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
-
S250A
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
-
S250C
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
-
S250T
-
the precursor is not processeed at the Gly249-Ser250 peptide bond. Production of an uncleaved and inactive protein
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
a validated method for measuring serum carnosinase activity in serum and heparin plasma
medicine
synthesis
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